Omer Fatima Abdelmutaal Ahmed, Hashim Najihah Binti Mohd, Ibrahim Mohamed Yousif, Dehghan Firouzeh, Yahayu Maizatulakmal, Karimian Hamed, Salim Landa Zeenelabdin Ali, Mohan Syam
1 Department of Pharmacy, Faculty of Medicine, University of Malaya, Kuala Lumpur, Malaysia.
2 Center for Natural Products Research and Drug Discovery, University of Malaya, Kuala Lumpur, Malaysia.
Tumour Biol. 2017 Nov;39(11):1010428317731451. doi: 10.1177/1010428317731451.
Xanthones are phytochemical compounds found in a number of fruits and vegetables. Characteristically, they are noted to be made of diverse properties based on their biological, biochemical, and pharmacological actions. Accordingly, the apoptosis mechanisms induced by beta-mangostin, a xanthone compound isolated from Cratoxylum arborescens in the human promyelocytic leukemia cell line (HL60) in vitro, were examined in this study. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay was done to estimate the cytotoxicity effect of β-mangostin on the HL60 cell line. Acridine orange/propidium iodide and Hoechst 33342 dyes and Annexin V tests were conducted to detect the apoptosis features. Caspase-3 and caspase-9 activities; reactive oxygen species; real-time polymerase chain reaction for Bcl-2, Bax, caspase-3, and caspase-9 Hsp70 genes; and western blot for p53, cytochrome c, and pro- and cleavage-caspase-3 and caspase-9 were assessed to examine the apoptosis mechanism. Cell-cycle analysis conducted revealed that β-mangostin inhibited the growth of HL60 at 58 µM in 24 h. The administration of β-mangostin with HL60 caused cell morphological changes related to apoptosis which increased the number of early and late apoptotic cells. The β-mangostin-catalyzed apoptosis action through caspase-3, caspase-7, and caspase-9 activation overproduced reactive oxygen species which downregulated the expression of antiapoptotic genes Bcl-2 and HSP70. Conversely, the expression of the apoptotic genes Bax, caspase-3, and caspase-9 were upregulated. Meanwhile, at the protein level, β-mangostin activated the formation of cleaved caspase-3 and caspase-9 and also upregulated the p53. β-mangostin arrested the cell cycle at the G/G phase. Overall, the results for β-mangostin showed an antiproliferative effect in HL60 via stopping the cell cycle at the G/G phase and prompted the intrinsic apoptosis pathway.
氧杂蒽酮是在多种水果和蔬菜中发现的植物化学化合物。其特点是，基于它们的生物学、生物化学和药理作用，具有多种特性。因此，本研究检测了从乔木黄牛木中分离出的一种氧杂蒽酮化合物β-山竹黄酮在体外对人早幼粒细胞白血病细胞系（HL60）诱导的凋亡机制。采用3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐法评估β-山竹黄酮对HL60细胞系的细胞毒性作用。进行吖啶橙/碘化丙啶和Hoechst 33342染色以及膜联蛋白V检测以检测凋亡特征。评估半胱天冬酶-3和半胱天冬酶-9活性；活性氧；Bcl-2、Bax、半胱天冬酶-3和半胱天冬酶-9、Hsp70基因的实时聚合酶链反应；以及p53、细胞色素c、前体和裂解的半胱天冬酶-3和半胱天冬酶-9的蛋白质印迹，以研究凋亡机制。进行的细胞周期分析显示，β-山竹黄酮在24小时内以58µM的浓度抑制HL60的生长。β-山竹黄酮与HL60共同作用导致与凋亡相关的细胞形态变化，增加了早期和晚期凋亡细胞的数量。β-山竹黄酮通过半胱天冬酶-3、半胱天冬酶-7和半胱天冬酶-9的激活催化凋亡作用，产生过量的活性氧，从而下调抗凋亡基因Bcl-2和HSP70的表达。相反，凋亡基因Bax、半胱天冬酶-3和半胱天冬酶-9的表达上调。同时，在蛋白质水平上，β-山竹黄酮激活裂解的半胱天冬酶-3和半胱天冬酶-9的形成，并上调p53。β-山竹黄酮使细胞周期停滞在G/G期。总体而言，β-山竹黄酮的结果显示其通过使细胞周期停滞在G/G期对HL60具有抗增殖作用，并促使内源性凋亡途径的发生。
